Antiviral amine derivatives of propanediols

ABSTRACT

Novel amine and amidine derivatives of di-O-(n-higher alkyl and alkenyl)-glycerols and -propanediols, and their pharmaceutically acceptable acid addition salts, are useful for combating viral infections in mammals. Of particular interest is 1,3-di-O-(n-hexadecyl)-2-O-(3-aminopropyl)-glycerol, and its pharmaceutically acceptable acid addition salts.

CROSS-REFERENCE TO RELATED APPLICATION

This application is a division of application Ser. No. 825,535 filedAug. 18, 1977.

BACKGROUND OF THE INVENTION

Virus infections which attack mammals, including man, are normallycontagious afflictions which are capable of causing great humansuffering and economic loss. Unfortunately, the discovery of antiviralcompounds is far more complicated and difficult than the discovery ofantibacterial and antifungal agents. This is due, in part, to the closestructural similarity of viruses and the structure of certain essentialcellular components such as ribonucleic and deoxyribonucleic acids.Nevertheless, numerous non-viral "antiviral agents", i.e. substances"which can produce either a protective or therapeutic effect to theclear detectable advantage of the virus infected host, or any materialthat can significantly enhance antibody formation, improve antibodyactivity improve non-specific resistance, speed convalescence or depresssymptoms" [Herrman et al., Proc. Soc. Exptl. Biol. Med., 103, 625(1960)], have been described in the literature. The list of reportedantiviral agents includes, to name a few, interferon and syntheticmaterials such as amantadine hydrochloride, pyrimidines, biguanides,guanidine, pteridines and methisazone. Because of the rather narrowrange of viral infections that can be treated by each of the antiviralagents commercially available at the present time, new syntheticantiviral agents are always welcomed as potentially valuable additionsto the armamentarium of medical technology.

The cells of mammals produce, in response to virus infection, asubstance which enables cells to resist the multiplication of a varietyof viruses. The viral-resisting or viral-interfering substances arereferred to as "interferons". The interferons are glycoproteins whichmay differ in their physico-chemical properties, but all exhibit thesame biological properties; namely, they inhibit a wide range ofunrelated viruses, have no toxic or other deleterious effects on cells,and are species-specific (Lockart, Frontiers of Biology, Vol. 2,"Interferons", edited by Finter, W. B. Saunders Co., Philadephia, 1966,pages 19-20).

No practical, economical method has yet been developed for thepreparation of exogenous interferon for routine clinical use againstviral infections. An alternative approach to producing interferon has,therefore, been pursued, which comprises administering to the animal tobe protected or treated a non-viral substance which stimulates--orinduces--production of interferon in the cells. The interferon producedin this fashion is referred to as "endogenous" interferon.

U.S. Pat. No. 2,738,351 discloses that compounds of the general formula##STR1## wherein each of R₁ and R₂ may be alkyl, aralkyl, aryl,cycloalkyl, nitro-substituted aryl, halogen-substituted aryl,alkyl-substituted aryl, or alkoxy-substituted aryl, each of X, Y and Zmay be oxygen, sulfur or sulfonyl, ALK is straight or branched alkyleneof from one to six carbon atoms, and B may be di(lower)alkylamino,piperidino, morpholino, pyrrolidino, (lower alkyl)pyrrolidino,N'-alkyl-piperazino or pipecolino, are local anesthetic agents.Additionally, the discussion of alternate synthetic routes (see Col. 1,11. 57-70, of said patent) discloses intermediates of the above formulawherein B is amino and (lower alkyl)amino. However, none of thecompounds specifically enumerate in the disclosure of said patentcontain an alkyl R₁ or R₂ larger than n-pentyl. Furthermore, in none ofthese compounds are both R₁ and R₂ alkyl and both X and Y oxygen.

Insecticidal and miticidal compounds of the formula ##STR2## wherein R₁and R₂ may each be, inter alia, lower alkylthio; q is 0 to 5; and A maybe, inter alia, 1-piperidino or di(lower alkyl)amino are disclosed inJapanese Pat. No. J7-6042-177.

SUMMARY OF THE INVENTION

It has now been discovered that certain novel amine and amidinederivatives of di-O-(n-higher alkyl and alkenyl)-glycerols and-propanediols are capable of combating viral infections in mammals. Thenovel compounds of this invention have the formulae ##STR3## and thepharmaceutically acceptable acid addition salts thereof, wherein

R₁ and R₂ are each selected from the group consisting of normal alkyl offrom 12 to 20 carbon atoms and normal alkenyl not having a double bondin the 1-position of from 12 to 20 carbon atoms,

Y is selected from the group consisting of alkylene of from 2 to 4carbon atoms, the two valencies being on different carbon atoms;##STR4## ortho-, meta- and para-phenylenedimethylene; ##STR5## wherein qis an integer of from one to three, and the left bond is connected to O;and ##STR6## wherein the left bond is connected to O,

Z is selected from the group consisting of alkylene of from 2 to 4carbon atoms, the two valencies being on different carbon atoms; ortho-,meta- and para-phenylenedimethylene; ##STR7## wherein q is an integer offrom one to three and the --(CH₂)_(q) -- group is connected to--(CH₂)_(p) NHR₃ ; an ##STR8## wherein the ##STR9## group is connectedto --(CH₂)_(p) NHR₃,

R₃ is selected from the group consisting of hydrogen, alkyl of from 2 to4 carbon atoms and ω-hydroxy(normal alkyl) of from 2 to 4 carbon atoms,

m, n and p are each 0 or 1, the sum of m, n and p being 0 or 1, R₃ beinghydrogen when m is 0, and R₃ being other than ω-hydroxy(normal alkyl)when m is 1 and Y is ##STR10##

W is selected from the group consisting of alkylene of from 1 to 4carbon atoms, the two valencies being on different carbon atoms when Wis other than methylene; ortho-, meta- and para-phenylene; and ##STR11##wherein the left bond is connected to O.

The invention disclosed herein comprises the novel antiviral compoundsof formulae I to V, the novel pharmaceutical compositions containing anantivirally effective amount of a compound of formulae I to V as theessential active ingredient in a pharmaceutically acceptable carrier,the novel method of prophylactically controlling a viral infection in amammal which comprises administering an amount effective toprophylactically control said viral infection of a compound of formulaeI to V, and the novel method of inducing the production of interferon ina mammal which comprises administering an amount effective to induce theproductio of interferon of a compound of formulae I to V.

DETAILED DESCRIPTION OF THE INVENTION

The compounds of this invention exhibit antiviral activity against awide variety of viruses in vivo in mammals and in vitro in mammaliantissue culture. At least a substantial portion of this activity resultsfrom the ability of said compounds to induce the production ofinterferon in the cells, i.e. endogenous interferon.

By "pharmaceutically acceptable" acid addition salts is meant thosesalts which are non-toxic at the dosages administered. Thepharmaceutically acceptable acid addition salts which may be employedinclude such water-soluble and water-insoluble salts as thehydrochloride, hydrobromide, phosphate, nitrate, sulfate, acetate,hexafluorophosphate, citrate, gluconate, benzoate, propionate, butyrate,sulfosalicylate, maleate, laurate, malate, fumarate, succinate, oxalate,tartrate, amsonate (4,4'-diaminostilbene-2,2'-disulfonate), pamoate(1,1'-methylene-bis-2-hydroxy-3-naphthoate), stearate,3-hydroxy-2-naphthoate, p-toluenesulfonate, methanesulfonate, lactate,and suramin salts.

One preferred group of the compounds of formulae I-V consists of thehydrochloride salts of the bases of formulae I-V.

Another preferred group of the compounds of formulae I-V consists ofthose wherein R₁ and R₂ are each normal alkyl of from 14 to 18 carbonatoms.

Another preferred group of the compounds of formulae I-V consists ofthose wherein R₁ and R₂ are each normal alkyl of from 14 to 18 carbonatoms and contain the same number of carbon atoms.

Another preferred group of the compounds of formulae I-V consists ofthose wherein R₁ and R₂ are each n-hexadecyl.

Another preferred group of the compounds of this invention consists ofthose of formula I.

Another preferred group of the compounds of this invention consists ofthose of formula II.

Another preferred group of the compounds of this invention consists ofthose of formula V.

One preferred group of the compounds of formulae I and II consists ofthose wherein m is 1, n is 0, p is 0, and R₃ is hydrogen.

Another preferred group of the compounds of formulae I and II consistsof those wherein m is 1, n is 0, p is 0, R₃ is hydrogen, and Y isstraight chain alkylene of from 2 to 4 carbon atoms.

Another preferred group of the compounds of formulae I and II consistsof those wherein m is 1, n is 0, p is 0, R₃ is hydrogen, and Y isortho-, meta- or para-phenylenedimethylene.

Particularly valuable are the following compounds and theirpharmaceutically acceptable acid addition salts:

1,3-di-O-(n-hexadecyl)-2-O-(3-aminopropyl)-glycerol,

1,2-di-O-(n-hexadecyl)-3-O-(3-aminopropyl)-glycerol,

1,3-di-O-(n-hexadecyl)-2-O-(meta-aminomethylbenzyl)-glycerol,

1,2-di-O-(n-hexadecyl)-3-O-(meta-aminomethylbenzyl)-glycerol,

1,2-di-O-(n-tetradecyl)-3-O-(meta-aminomethylbenzyl)-glycerol

1,3-di-O-(n-hexadecyl)-2-O-(meta-aminomethylphenyl)-glycerol,

1,3-di-O-(n-hexadecyl)-2-O-(para-aminomethylphenyl)-glycerol,

1,2-di-O-(n-hexadecyl)-3-O-(para-aminomethylphenyl)-glycerol,

1,2-di(n-hexadecyloxy)-3-(meta-aminomethylbenzylamino)-propane,

1,2-di(n-hexadecyloxy)-3-aminomethyl-propane,

1,2-di-O-(n-hexadecyl)-3-O-(meta-amidinobenzyl)-glycerol,

1-[2,3-di(n-octadecyloxy)propyl]-4-aminomethyl-4-phenylpiperidine,

1-[2,3-di(n-hexadecyloxy)propyl]-4-aminomethyl-4-phenylpiperidine, and

1-[2,3-di(n-tetradecyloxy)propyl]-4-aminomethyl-4-phenylpiperidine.

The compounds of formulae I and II above may be prepared from theappropriate 1,2-di-O-(n-higher alkyl or alkenyl)-glycerol and1,3-di-O-(n-higher alkyl or alkenyl)-glycerol starting materials bymethods familiar to those skilled in the art. For example:

(a) those compounds wherein m is 1, R₃ is H and Y is 3-propylene may beprepared by condensing the starting material with acrylonitrile inaqueous solution under strongly basic conditions to form the2-cyanoethyl derivative, which is then hydrogenated;

(b) those compounds wherein m is 1, R₃ is H and Y is 2-ethylene may beprepared by reacting the 2-cyanoethyl derivative of the startingmaterial with formic acid under strongly acidic conditions to form the2-carboxyethyl derivative, which is then reacted under strongly acidicconditions with hydrazoic acid;

(c) those compounds wherein m is 1, R₃ is H and Y is 4-butylene may beprepared by adding an allyl radical to the starting material by reactingit with an allyl halide under strongly basic conditions, hydroboratingthe allyl derivative, oxidizing the resulting intermediate with hydrogenperoxide in basic aqueous solution to the 3-hydroxypropyl derivative,reacting the 3-hydroxypropyl derivative with a sulfonyl chloride RSO₂ Cl(e.g. p-toluenesulfonyl chloride) under basic conditions to form thecorresponding sulfonate ester (e.g. the tosylate), substituting a cyanogroup for the RSO₃ - group by reaction with sodium cyanide, and thenreducing the resulting 3-cyanopropyl derivative;

(d) those compounds wherein m is 1, R₃ is H and Y is 2-propylene may beprepared by following procedure (c) through the hydrogen peroxideoxidation step, isolating the 2-hydroxypropyl oxidation side product,and then subjecting the 2-hydroxypropyl derivative to the remainder ofprocedure (c) using sodium azide in place of sodium cyanide;

(e) those compounds wherein m is 1, R₃ is H and Y is2-hydroxy-3-propylene may be prepared by oxidizing the allyl derivativeof the starting material with a percarboxylic acid (e.g.m-chloroperbenzoic acid) to the 2,3-epoxypropyl derivative, and reactingthe latter with sodium azide to form the 3-azido-2-hydroxypropylderivative, which is then reduced;

(f) those compounds wherein m is 1, R₃ is H and Y isphenylenedimethylene may be prepared by reacting the starting materialwith a cyanobenzyl halide under strongly basic conditions, and thenreducing the resulting cyanobenzyl derivative with a hydride reagentsuch as lithium aluminum hydride;

(g) those compounds wherein m is 1, R₃ is H, Y is ##STR12## and q is aninteger of from one to three may be prepared by reacting the startingmaterial with a sulfonyl chloride RSO₂ Cl (e.g. p-toluenesulfonylchloride) under basic conditions to form the corresponding sulfonateester (e.g. the tosylate) of di-O-(n-higher alkyl or alkenyl)-glycerol,substituting a cyanophenoxy or ω-cyanoalkylphenoxy group for the RSO₃ --group by reaction with e.g. sodium cyanophenolate and then hydrogenatingthe resulting cyanophenyl or cyanoalkylphenyl derivative of thedi-O-(n-higher alkyl or alkenyl)-glycerol starting material;

(h) those compounds wherein m is 1, R₃ is normal alkyl and Y is otherthan ##STR13## may be prepared by acylating the corresponding compoundwherein R₃ is H with an acyl halide under basic conditions, and thenreducing the resulting N-acyl derivative;

(i) those compounds wherein m is 1, R₃ is isopropyl and Y is other than##STR14## may be prepared by reacting the corresponding compound whereinR₃ is H with acetone under acidic conditions and hydrogenating theresulting imine (e.g. with sodium borohydride);

(j) those compounds wherein m is 1, R₃ is other than H and Y is2-ethylene may be prepared by oxidizing the allyl derivative of thedi-O-(n-higher alkyl or alkenyl)-glycerol starting material bysequential treatment in the presence of water with osmium tetroxide (orpotassium permanganate) and sodium periodate to the formylmethylderivative, reacting the formylmethyl derivative with the amine R₃ NH₂under acidic conditions, and hydrogenating the N-alkylidene orN-hydroxyalkylidene product;

(k) those compounds wherein m is 1, R₃ is alkyl and Y is ##STR15## maybe prepared by reacting the 2,3-epoxypropyl derivative of the startingmaterial with the amine R₃ NH₂ ;

(l) those compounds wherein p is 1 may be prepared by reacting asulfonate ester (e.g. the tosylate) of the appropriate di-O-(n-higheralkyl or alkenyl)-glycerol [prepared from the starting material as in(g)] with sodium cyanide, and then hydrogenating the resulting cyanoderivative of di(n-higher alkyloxy or alkenyloxy)propane;

(m) those compounds wherein m, n and p are all 0 may be prepared as in(1) using sodium azide in place of sodium cyanide;

(n) those compounds wherein n is 1 and Z is 3-propylene may be preparedby condensing the corresponding compound wherein m, n and p are all 0with acrylonitrile in aqueous solution under strongly basic conditionsto form the N-(2-cyanoethyl)-amino derivative of di(n-higher alkyloxy oralkenyloxy)propane, which is then hydrogenated;

(o) those compounds wherein n is 1 and Z is phenylenedimethylene may beprepared by reacting a xylylenediamine with a sulfonate ester (e.g. thetosylate) of the appropriate di-O-(n-higher alkyl or alkenyl)-glycerol;and

(p) those compounds wherein m is 1, R₃ is alkyl and Y is ##STR16##

may be prepared by reducing a cyanobenzyl derivative of thedi-O-(n-higher alkyl or alkenyl)-glycerol starting material to theformylbenzyl derivative, reducing the formylbenzyl derivative in thepresence of trimethylsulfonium iodide to the 1,2-epoxyethylbenzylderivative, and then reacting the latter derivative with the amine R₃NH₂.

The skilled worker in the art will realize that additional compounds offormulae I and II may be prepared by using obvious variations of themethods of synthesis outlined above.

The compounds of formulae III and IV above may also be prepare from theappropriate 1,2-di-O-(n-higher alkyl or alkenyl)-glycerol and1,3-di-O-(n-higher alkyl or alkenyl)-glycerol starting material bymethods familiar to those skilled in the art. For example, thosecompounds wherein W is phenylene may be prepared by condensing acyanophenyl derivative of the starting material with ethanol orethanethiol in a hydrogen chloride saturated inert solvent such asdioxane to form the corresponding ethylbenzimidate orethylthiobenzimidate hydrochloride, followed by nucleophilicsubstitution with ammonia and elimination of ethanol or ethanethiolwhich is carried out in ammonia saturated ethanol. Those compoundswherein W is ##STR17## may be prepared in like manner from a cyanobenzylderivative of the starting material. Those compounds where W is alkylenemay be prepared in like manner from the appropriate cyano (lower alkyl)derivative of the starting material. When W is methylene, saidderivative may be prepared by reacting the starting material withchloro-, bromo-, or iodoacetonitrile.

The compounds of formula V above may be prepared from the appropriate1,2-di-O-(n-higher alkyl or alkenyl)-glycerol starting materials bymethods familiar to those skilled in the art. For example, the tosylderivative of the starting material may be reacted with4-cyano-4-phenylpiperidine hydrochloride, and the resulting compoundthen reduced.

Acid addition salts of the bases of formulae I-V may be prepared byconventional procedures such as by mixing the amine or amidine compoundin a suitable solvent with the required acid and recovering the salt byevaporation or by precipitation upon adding a non-solvent for the salt.Hydrochloride salts may readily be prepared by passing hydrogen chloridethrough a solution of the amine or amidine compound in an organicsolvent. As can be seen by reference to the examples herein, many of theisolated hydrochloride or dihydrochloride salts of the bases of formulaeI-V tend to contain a significant water content. Whether this observed"trapped" water is randomly occluded during crystallization, orcorresponds to formation of true molecular hydrates, or results from theoccurrence of some other phenomenon, is not known. In any event, thesalts containing "trapped" water may be efficaciously formulated andadministered without preliminary dehydration.

The 1,2-di-O-(n-higher alkyl)-glycerol starting materials may beprepared by the method of Kates, M. et al., Biochemistry, 2, 394 (1963).The 1,3-di-O-(n-higher alkyl)-glycerol starting materials may beprepared by the method of Damico, R., et al., J. Lipid Res., 8, 63(1967). The 1,2- and 1,3-di-O-(n-higher alkenyl)-glycerol startingmaterials may be prepared by the method of Bauman, W. J. and Mangold, H.K., J. Org. Chem., 31, 498 (1966).

The antiviral activity of the compounds of this invention was determinedby the use of two independent procedures. In the first, the testcompound is administered to mice by the intraperitoneal route eighteento twenty-four hours prior to challenging them with a lethal dose ofencephalomyocarditis (EMC) virus. Survival data are taken during the tendays after challenge and compared with the data for unprotected animals.The procedure in which the drug is given eighteen to twenty-four hoursbefore, and at a distinctly different site from, virus injection isdesigned to eliminate local effects between drug and virus and identifyonly those compounds which produce a systemic antiviral response.

In the second procedure, monolayers of human nasal polyp cells grown onmicrotiter plates are treated with the test compound about eighteenhours before treatment with a lethal dose of vesicular stomatitis virus(VSV). The test compound is washed away from the monolayers before virustreatment. Culture fluid extracted from the plates after a postchallenge incubation period is titrated for the amount of infectiousvirus present in microtiter plates of L-929 mouse fibroblasts.Comparison is made with the virus yield data for culture fluid extractedfrom unprotected polyp cells.

Additionally, many of the compounds of this invention were tested fortheir ability to enhance the known antiviral activity ofpolyinosinic:polycytidylic acid. Finally, certain of the compounds werealso tested for their ability to induce circulating interferon in miceafter parenteral administration, using the procedure described byHoffman, W. W., et al., Antimicrobial Agents and Chemotherapy, 3,498-501 (1973).

Parenteral, topical or intranasal administration of the above-describedamines and amidines to a mammal before exposure of the mammal to aninfectious virus provides rapid resistance to the virus. Preferably,administration should take place from about two days to about one daybefore exposure to the virus, although this will vary somewhat with theparticular animal species and the particular infectious virus.

When the materials of this invention are administered, they are mosteasily and economicaly used in a dispersed form in an acceptablecarrier. When it is said that this material is dispersed, it means thatthe particles may be molecular in size and held in true solution in asuitable solvent or that the particles may be colloidal in size anddispersed through a liquid phase in the form of a suspension or anemulsion. The term "dispersed" also means that the particles may bemixed with and spread throughout a solid carrier so that the mixture isin the form of a powder or dust. This term is also meant to encompassmixtures which are suitable for use as sprays, including solutions,suspensions or emulsions of the agents of this invention.

When administered parenterally (subcutaneously, intramuscularly,intraperitoneally) the materials of this invention are used at a levelof from about 1 mg./kg. of body weight to about 250 mg./kg. body weight.The favored range is from about 5 mg./kg. to about 100 mg./kg. of bodyweight, and the preferred range from about 5 mg. to about 50 mg./kg. ofbody weight. The dosage, of course, is dependent upon the mammal beingtreated and the particular amine or amidine compound involved and is tobe determined by the individual responsible for its administration.Generally, small doses will be administered initially with gradualincrease in dosage until the optimal dosage level is determined for theparticular subject under treatment.

Vehicles suitable for parenteral injection may be either aqueous such aswater, isotonic saline, isotonic dextrose, Ringer's solution, ornon-aqueous such as fatty oils of vegetable origin (cottonseed, peanutoil, corn, sesame) and other non-aqueous vehicles which will notinterfere with the efficacy of the preparation and are non-toxic in thevolume or proportion used (glycerol, ethanol, propylene glycol,sorbitol). Additionally, compositions suitable for extemporaneouspreparation of solutions prior to administration may advantageously bemade. Such compositions may include liquid diluents, for example,propylene glycol, diethyl carbonate, glycerol, sorbitol.

In practicing the intranasal route of administration of this inventionany practical method can be used to contact the antiviral agent with therespiratory tract of the mammal. Effective methods includeadministration of the agent by intranasal or nasopharyngeal drops and byinhalation as delivered by a nebulizer or an aerosol. Such methods ofadministration are of practical importance because they provide an easy,safe and efficient method of practicing this invention. For intranasaladministration of the agent, usually in an acceptable carrier, aconcentration of agent between 1.0 mg./ml. and 100 mg./ml. issatisfactory. Concentrations in the range of about 30 to 50 mg./ml.allow administration of a convenient volume of material.

For topical application the antiviral agents are most conveniently usedin an acceptable carrier to permit ease and control of application andbetter absorption. Here also concentrations in the range of from about1.0 mg./ml. to about 250 mg./ml. are satisfactory. In general, in theabove two methods of administration a dose within the range of about 1.0mg./kg. to about 250 mg./kg. of body weight and, preferably, from about5.0 mg./kg. to about 50 mg./kg. of body weight will be administered.

The compounds employed in this invention may be employed alone, i.e.,without other medicinals, as mixtures of more than one of theherein-described compounds, or in combination with other medicinalagents, such as analgesics, anesthetics, antiseptics, decongestants,antbiotics, vaccines, buffering agents and inorganic salts, to afforddesirable pharmacological properties. Further, they may be administeredin combination with hyaluronidase to avoid or, at least, to minimizelocal irritation and to increase the rate of absorption of the compound.Hyaluronidase levels of at least about 150 (U.S.P.) units are effectivein this respect although higher or lower levels can, of course, be used.

Those materials of this invention which are water-insoluble, includingthose which are of low and/or difficult solubility in water, are, foroptimum results, administered in formulations, e.g., suspensions,emulsions, which permit formation of particle sizes of less than about20μ. The particle sizes of the formulations influence their biologicalactivity apparently through better absorption of the active materials.In formulating these materials various surface active agents andprotective colloids are used. Suitable surface active agents are thepartial esters of common fatty acids, such as lauric, oleic, stearic,with hexitol anhydrides derived from sorbitol, and the polyoxyethylenederivatives of such ester products. Such products are sold under thetrademarks "Spans" and "Tweens," respectively, and are available fromICI United States Inc., Wilmington, Del. Cellulose ethers, especiallycellulose methyl ether (Methocel, available from the Dow Chemical Co.,Midland, Mich.) are highly efficient as protective colloids for use inemulsions containing the materials of this invention.

The water-soluble materials described herein are administered foroptimum results in aqueous solution. Typically they are administered inphosphate buffered saline. The water-insoluble compounds areadministered in formulations of the type described above or in variousother formulations as previously noted. Dimethylsulfoxide serves as asuitable vehicle for water-insoluble compounds. A representativeformulation for such compounds comprises formulating 25 to 100 mg. ofthe chosen drug as an emulsion by melting and mixing with equal parts ofpolysorbate 80 and glycerin to which hot (80° C.) water is added undervigorous mixing. Sodium chloride is added in a concentration solution toa final concentration of 0.14 M and sodium phosphate, pH 7, is added toa final concentration of 0.01 M to give, for example, the followingrepresentative composition:

    ______________________________________                                                               mg./ml.                                                ______________________________________                                        Drug                     50.0                                                 Polysorbate 80           50.0                                                 Glycerin                 50.0                                                 Sodium Phosphate Monobasic Hydrous                                                                     1.4                                                  Sodium Chloride          7.9                                                  Water                    842.0                                                                         1001.3                                               ______________________________________                                    

In certain instances, as where clumping of the drug particles occurs,sonication is employed to provide a homogenous system.

The following examples illustrate the invention but are not to beconstrued as limiting the same.

EXAMPLE 1 1,3-Di-O-(n-hexadecyl)-2-O-(3-aminopropyl)-glycerolHydrochloride A. 1,3-Di-O-(n-hexadecyl)-2-O-(2-cyanoethyl)-glycerol

A mixture of 1,3-di-O-(n-hexadecyl)-glycerol (80 g., 148 mmoles),acrylonitrile (1.49 kg., 28.1 moles) and aqueous 2 N sodium hydroxide(1.2 l.) was heated to 50° C. Tetrabutylammonium hydroxide (19.2 g. of40 wt. % aqueous solution, 29.15 mmoles) was slowly added, causing thetemperature of the exothermic reaction mixture to rise to about 80° to90° C. The reaction mixture was then stirred for 20 minutes without anyexternal heating, followed by cooling to 20° C. and addition of water(1.0 l.). A solid material, a mixture of unreacted and cyanoethylated1,3-di-O-(n-hexadecyl)-glycerol, was isolated and treated again withfresh acrylonitrile (1.49 kg., 28.1 moles), aqueous 2 N sodium hydroxide(1.2 l.) and tetrabutylammonium hydroxide (19.2 g. of 40 wt. % aqueoussolution, 29.15 mmoles) for 20 minutes with stirring at 50° C., followedby cooling and addition of water (1.0 l.). The resulting1,3-di-O-(n-hexadecyl)-2-O-(2-cyanoethyl)-glycerol solids were filtered,washed consecutively with water, methanol and acetonitrile, and dried[82 g., 93% yield, m.p. 45°-46° C., ir (CHCl₃) 2250 cm⁻¹, n.m.r. (CDCl₃)δ 3.92 (t, 2, NCCH₂ CH₂ O--), 3.33-3.67 (m, 9, --OCH[CH₂ OCH₂ C₁₅ H₃₁]₂), 2.62 (t, 2, NCCH₂ CH₂ O--) and 0.75-1.58 (m, 62, aliphaticprotons)].

B. Title Compound

A mixture of 1,3-di-O-(n-hexadecyl)-2-O-(2-cyanomethyl)-glycerol (20.5g., 34.5 mmoles), tetrahydrofuran (200 ml.), ethanol (10 ml.) and Raneynickel catalyst (3 g.) was saturated with ammonia gas at 0° to 5° C. andthen hydrogenated (50 psi) in a Paar hydrogenator for 3 hours at roomtemperature. The mixture was then filtered, the catalyst washed withtetrahydrofuran (50 ml.), and the total filtrate evaporated in vacuo toan oil. This procedure was repeated three more times with freshreactants and catalyst to yield a total of 77 g. of oil. The oil wasdissolved in ether (500 ml.) and the solution washed with 2 wt. %aqueous ammonium hydroxide solution (500 ml.), dried (MgSO₄), filteredand evaporated in vacuo to yield a solid. The solid was dissolved inmethanol (300 ml.) and the solution saturated with hydrogen chloride gasand then evaporated in vacuo to a solid. This solid was crystallizedfrom ethyl acetate to yield the named product with a slight impurity (63g., 72% yield, m.p. 69°-70° C.), and then recrystallized twice fromisopropanol:acetonitrile (1:1, 800 ml.) [47.5 g., 54% yield, m.p.58°-59° C., n.m.r. (CDCl₃) δ 3.84 (t, 2, H₂ NCH₂ CH₂ CH₂ O--), 3.55 (m,9, --OCH[CH₂ OCH₂ C₁₅ H₃₁ ]₂), 3.24 (t, 2, H₂ NCH₂ CH₂ CH₂ O--), 2.04(m, 2, H₂ NCH₂ CH₂ CH₂ O--) and 0.90-1.32 (m, 62, aliphatic protons),elemental analysis calculated: 72.04% C; 12.73% H; 2.21% N; found:71.80% C; 12.41% H; 2.30% N].

EXAMPLES 2-7

In like manner to that described in Example 1 the following compoundswere prepared by using the appropriate 1,3- or 1,2-di-O-(n-higheralkyl)-glycerol as starting material:

    __________________________________________________________________________     ##STR18##                                                   I                 ##STR19##                                                   II                                                            Elemental Analysis               Example                   Molecular          Calculated                                                                             Found (%)               Number                                                                             Structure                                                                          R.sub.1 R.sub.2 Formula      M.P. (°C.)                                                                   C  H  N  C  H  N                 __________________________________________________________________________    2    I    n-dodecyl                                                                             n-dodecyl                                                                             C.sub.30 H.sub.63 O.sub.3 N . HCl . 3/2 H.sub.2                               O            76-77 65.60                                                                            12.29                                                                            2.54                                                                             65.45                                                                            11.91                                                                            2.61              3    I    n-tetradecyl                                                                          n-tetradecyl                                                                          C.sub.34 H.sub.71 O.sub.3 N . HCl . H.sub.2                                                57-58 68.47                                                                            12.50                                                                            2.35                                                                             68.51                                                                            11.24                                                                            2.29              4    I    n-octadecyl                                                                           n-octadecyl                                                                           C.sub.42 H.sub.87 O.sub.3 N .                                                              64-65 73.04                                                                            12.84                                                                            2.03                                                                             72.96                                                                            12.56                                                                            1.99              5    II   n-tetradecyl                                                                          n-tetradecyl                                                                          C.sub.34 H.sub.71 O.sub.3 N .                                                              90-91 70.60                                                                            12.55                                                                            2.42                                                                             70.74                                                                            12.85                                                                            2.68              6    II   n-hexadecyl                                                                           n-hexadecyl                                                                           C.sub.38 H.sub.79 O.sub.3 N . HCl . 3/4 H.sub.2                               O            76-78 70.54                                                                            12.77                                                                            2.16                                                                             70.42                                                                            12.17                                                                            2.07              7    II   n-octadecyl                                                                           n-octadecyl                                                                           C.sub.42 H.sub.87 O.sub.3 N . HCl . H.sub.2                                                67-69 71.19                                                                            12.80                                                                            1.98                                                                             70.95                                                                            12.19                                                                            1.91              __________________________________________________________________________

EXAMPLE 8 1,3-Di-O-(n-hexadecyl)-2-O-(2-aminoethyl)-glycerolHydrochloride A. 1,3-di-O-(n-hexadecyl)-2-O-(2-carboxyethyl)-glycerol

A mixture of 1,3-di-O-(n-hexadecyl)-2-O-(2-cyanoethyl)-glycerol (4.8 g.,8.1 mmoles), concentrated hydrochloric acid (50 ml.) and formic acid (50ml.) was stirred for 16 hours at reflux, then cooled and extracted withether (3×100 ml.). The combined ether extract was washed with water (200ml.), dried (MgSO₄), filtered and evaporated in vacuo to yield1,3-di-O-(n-hexadecyl)-2-O-(2-carboxyethyl)-glycerol solids (4.5 g.),which were purified by silica gel chromatography (elution withtoluene:ethanol) [3.5 g., 71% yield, m.p. 43°-45° C., ir (CHCl₃) 1740cm⁻¹, n.m.r. (CDCl₃) δ 3.93 (t, J=6 Hz, 2, --OCH₂ CH₂ COOH) and 2.65 (t,J=6 Hz, 2, --OCH₂ CH₂ COOH)].

B. Title Compound

1,3-Di-O-(n-hexadecyl)-2-O-(2-carboxyethyl)-glycerol (3.5 g., 5.7mmoles) was dissolved in a mixture of benzene (55 ml.) and concentratedsulfuric acid (5.89 g.). Hydrazoic acid (6.34 ml. of 4.65 wt. % benzenesolution, 6.0 mmoles) was then added dropwise and the resulting mixturestirred for 2 hours at room temperature. Thin layer chromatography (TLC)analysis showed about 50% reaction of the 2-carboxyethyl compound.Additional hydrazoic acid (6.34 ml. of 4.65 wt. % benzene solution, 6.0mmoles) was added dropwise and the reaction mixture stirred for another16 hours at 40° C. TLC analysis now showed that the reaction wasessentially complete. Water (50 ml.) and aqueous 2 N sodium hydroxidewere then added and the resulting mixture extracted with ether (3×200ml.). The combined ether extract was dried (Na₂ SO₄), filtered,saturated with hydrogen chloride gas and evaporated in vacuo to yield asolid. The solid was purified by silica gel chromatography (elution withchloroform:methanol) and recrystallized from hot ethyl acetate [570 mg.,16% yield, m.p. 79°-80° C., n.m.r. (CDCl₃) δ 3.95 (m, 2, --OCH₂ CH₂ NH₂)and 3.22 (m, 2, --OCH₂ CH₂ NH₂), elemental analysis calculated: 71.62%C; 12.67% H; 2.26% N; found: 70.90% C; 12.19% H; 2.05% N].

EXAMPLE 9 1,3-Di-O-(n-hexadecyl)-2-O-(3-ethylaminopropyl)-glycerolHydrochloride A. 1,3-Di-O-(n-hexadecyl)-2-O-(3-acetamidopropyl)-glycerol

1,3-Di-O-(n-hexadecyl)-2-O-(3-aminopropyl)-glycerol hydrochloride (1.0g., 1.6 mmoles) was added to a mixture of potassium carbonate (830 mg.,6.0 mmoles) and benzene (75 ml.). Acetyl chloride (150 mg., 1.9 mmoles)was then added and the resulting mixture stirred for one hour at reflux.Additional acetyl chloride (150 mg., 1.9 moles) was added and thereaction mixture stirred for another hour at reflux. TLC analysis showedthat the reaction was essentially complete. The reaction mixture wascooled, water (75 ml.) added, and the resulting mixture extracted withether (3×100 ml.). The combined ether extract was dried (MgSO₄),filtered and evaporated in vacuo to yield the named compound [800 mg.,79% yield, m.p. 53°-54° C., ir (CHCl₃) 3400 and 1670 cm⁻¹, n.m.r.(CDCl₃) δ 1.97 (s, 3, --NHCOCH₃)].

B. Title Compound

1,3-Di-O-(n-hexadecyl)-2-O-(3-acetamidopropyl)-glycerol (700 mg., 1.1mmoles) was dissolved in ether (100 ml.) and treated with lithiumaluminum hydride (500 mg., 13 mmoles). Water (100 ml.) was then addedand the mixture extracted with ether (2×100 ml.). The combined etherextract was dried (MgSO₄), filtered, treated with hydrogen chloride gasand evaporated in vacuo to a solid, which was recrystallized from hotethyl acetate [470 mg., 66% yield, m.p. 61°-62° C., n.m.r. (CDCl₃) δ1.47 (t, 3, --NHCH₂ CH₃), elemental analysis calculated: 72.51% C;12.78% H; 2.11% N; found: 72.47% C; 12.56% H; 2.03% N].

EXAMPLE 10 1,3-Di-O-(n-hexadecyl)-2-O-(3-isopropylaminopropyl)-glycerolHydrochloride

1,3-Di-O-(n-hexadecyl)-2-O-(3-aminopropyl)-glycerol hydrochloride (700mg., 1.1 mmoles) was dissolved in a solution of acetic acid (1.05 ml.),sodium acetate (350 mg., 4.3 mmoles) and acetone (1.3 ml.). Sodiumborohydride (1.25 g., 33 mmoles) was added in small portions until TLCanalysis showed that all the 3-aminopropyl compound had been consumed.The reaction mixture was then treated with aqueous 2 N sodium hydroxide(20 ml.) and water (20 ml.), and extracted with ether (3×40 ml.). Thecombined ether extract was dried (MgSO₄), filtered, treated withhydrogen chloride gas, and then evaporated in vacuo to a solid, whichwas recrystallized from hot ethyl acetate [210 mg., solid containedabout 1/2 mole H₂ O per mole named product, 28% yield, m.p. 72°-73° C.,n.m.r. (CDCl₃) δ 1.42 (d, 6, --NHCH[CH₃ ]₂), elemental analysiscalculated: 71.82% C; 12.79% H; 2.04% N; found: 71.92% C; 12.46% H;1.94% N].

EXAMPLE 11 1,2-Di-O-(n-hexadecyl)-3-O-(2-isopropylaminomethyl)-glycerolHydrochloride A. 1,2-Di-O-(n-hexadecyl)-3-O-allyl-glycerol

Sodium hydride (1.78 g. of 50 wt. % dispersion in mineral oil, 37mmoles) was added at 60° C. to a solution of1,2-di-O-(n-hexadecyl)-glycerol (10 g., 18.5 mmoles) inN,N-dimethylformamide (100 ml.), and the resulting solution stirred for20 minutes at 60° C. Allyl bromide (4.47 g., 37 mmoles) was then addeddropwise and the resulting mixture stirred for 3 hours at 90° C.,cooled, cautiosly diluted with water (200 ml.) to quench the reaction,and extracted with ether (3×150 ml.). The combined ether extract waswashed with saturated aqueous sodium chloride solution, dried (MgSO₄),filtered and evaporated in vacuo to an oil, which was purified by silicagel chromatography (elution with benzene) [10 g., 93% yield, oil, n.m.r.(CDCl₃) δ 5.66-6.16 (m, 1, --OCH₂ CH═CH₂), 5.25 (d of doublets, 2,--OCH₂ CH═CH₂) and 4.03 (d, 2, --OCH₂ CH═CH₂)].

B. 1,2-Di-O-(n-hexadecyl)-3-O-formylmethyl-glycerol

Osmium tetroxide (90 mg., 0.354 mmoles) was added to a solution of1,2-di-O-(n-hexadecyl)-3-O-allyl-glycerol (4.5 g., 7.75 mmoles) intetrahydrofuran:water (3:1, 120 ml.), and the resulting solution stirredfor 5 minutes at room temperature. Sodium periodate (9 g., 42 mmoles)was then added and the reaction solution stirred for 16 hours at roomtemperature under nitrogen. The reaction solution was then diluted withwater (150 ml.) and extracted with ether (2×150 ml.). The combined etherextract was washed with water (150 ml.), dried (MgSO₄) and evaporated invacuo to an oil, which was purified by silica gel chromatography(elution with benzene:ethyl acetate) [2.6 g., 57% yield, waxy solid, ir(CHCl₃) 1735 cm⁻¹, n.m.r. (CDCl₃) δ 9.38 (t, J=1 Hz, 1, --OCH₂ CHO) and4.07 (d, J=1 Hz, 2, --OCH₂ CHO)].

C. Title Compound

Sodium cyanoborohydride (0.1 g., 1.6 mmoles) was added to a solution of1,2-di-O-(n-hexadecyl)-3-O-formylmethyl-glycerol (1.5 g., 2.6 mmoles)and isopropylamine (0.89 g., 15 mmoles) in methanol:tetrahydrofuran(1:1, 50 ml.), and the mixture stirred for two hours at roomtemperature. The pH was then adjusted to 6 with 5 N methanolichydrochloric acid, additional sodium cyanoborohydride (0.1 g., 1.6mmoles) added, and the reaction mixture then stirred for another 60hours at room temperature, filtered, treated with aqueous 3 N sodiumhydroxide (10 ml.) and saturated aqueous sodium chloride solution (200ml.), and extracted with ether (2×150 ml.). The combined ether extractwas dried (MgSO₄), filtered and evaporated in vacuo to an oily solid,which was purified by silica gel chromatography (elution withbenzene:ethanol) and dissolved in methanol. The solution was treatedwith hydrogen chloride gas and evaporated in vacuo to yield a solid,which was recrystallized from ethyl acetate [400 mg., solid containedabout 1/4 mole H.sub. 2 O per mole named product, 23% yield, m.p.71°-72° C., n.m.r. (CDCl₃) δ 1.42 (d, J=6 Hz, 6, --NHCH[CH₃ ]₂),elemental analysis calculated: 72.02% C; 12.76% H; 2.10% N; found:71.89% C; 12.34% H; 2.09% N].

EXAMPLE 121,2-Di-O-(n-hexadecyl)-3-O-[2-(2-hydroxyethylamino)-ethyl]-glycerolHydrochloride

In like manner to that described in Example 11 the named compound wasprepared by reacting 2-hydroxyethylamine with1,2-di-O-(n-hexadecyl)-3-O-formylmethyl-glycerol [solid contained about1/2 mole H₂ O per mole named product, m.p. 125°-126° C., elementalanalysis calculated: 69.54% C; 12.42% H; 2.07% N; found: 69.62% C;12.08% H; 2.29% N].

EXAMPLE 13 1,3-Di-O-(n-hexadecyl)-2-O-(4-aminobutyl)-glycerolHydrochloride A. 1,3-Di-O-(n-hexadecyl)-2-O-(3-hydroxypropyl)-glycerol

Borane methyl sulfide (BMS) complex (6.5 ml., 68.5 mmoles) was added at0° to 5° C. to a solution of 1,3-di-O-(n-hexadecyl)-2-O-allyl-glycerol(10.82 g., 18.6 mmoles, prepared as in Example 11A) in hexane (190 ml.),and the resulting solution stirred for 3 hours at room temperature. Thereaction solution was then cooled again to 0° to 5° C. and ethanol (17.3ml.) added dropwise to decompose residual BMS. The reaction solution wasthen treated with aqueous 3 N sodium hydroxide (13 ml.) and 30 wt. %aqueous hydrogen peroxide (11 ml.), stirred for 16 hours at reflux,cooled, and poured into ice water containing sodium bisulfite. The icewater solution was stirred until it gave a negative starch-iodide testfor peroxides, and then extracted with ether (3×200 ml.). The combinedether extract was washed with water (200 ml.), washed with saturatedaqueous sodium chloride solution (200 ml.), dried (MgSO₄), filtered andevaporated in vacuo. The resulting product was purified by silica gelchromatography (elution with benzene:ethanol) [5 g., 45% yield, m.p. 29°C., n.m.r. (CDCl₃) δ 3.80 (t, J=5 Hz, 2, --OCH₂ CH₂ CH₂ OH) and 3.75 (t,J=5 Hz, 2, --OCH₂ CH₂ CH₂ OH)].

B. 1,3-Di-O-(n-hexadecyl)-2-O-[3-(p-tosyloxy)propyl]-glycerol

1,3-Di-O-(n-hexadecyl)-2-O-(3-hydryoxypropyl)-glycerol (8.0 g., 13.4mmoles) was added at 10° C. to a solution of p-toluenesulfonyl chloride(5.25 g., 27.5 mmoles) and pyridine (10 ml. in methylene chloride (200ml.), and the mixture stirred for 60 hours at room temperature. Water(200 ml.) was then added, the methylene chloride and aqueous phasesseparated, and the latter extracted with methylene chloride (2×150 ml.).The three methylene chloride layers were combined, washed with water(2×150 ml.), dried (MgSO₄), filtered and evaporated in vacuo. Theresulting tosylate was purified by silica gel chromatography (elutionwith benzene) [3.0 g., 30% yield, oil, ir (CHCl₃) 1130 and 1350 cm⁻¹,n.m.r. (CDCl₃) δ 7.53 (q, 4, protons on phenyl ring), 4.15 (t, 2, --SO₃CH₂ CH₂ CH₂ O--), 3.63 (t, 2, --SO₃ CH₂ CH₂ CH₂ O--), 3.42 (m, 9,--OCH[CH₂ OCH₂ C₁₅ H₃₁ ]₂), 2.45 L (s, 3, Ar--CH₃), 1.90 (m, 2, --SO₃CH₂ CH₂ CH₂ O--) and 0.90-1.50 (m, 62, aliphatic protons)].

C. 1,3-Di-O-(n-hexadecyl)- 2-O-(3-cyanopropyl)-glycerol

1,3-Di-O-(n-hexadecyl)-2-O-[3-(p-tosyloxy)propyl]-glycerol (3.0 g., 4.0mmoles) was dissolved in a solution of sodium cyanide (0.5 g., 10mmoles) in N,N-dimethylformamide (50 ml.), and the resulting solutionstirred for 16 hours at 80° C., cooled, diluted with water (100 ml.) andextracted with ether (3×100 ml.). The combined ether extract was washedconsecutively with 1 N hydrochloric acid (3×75 ml.), saturated aqueoussodium bicarbonate solution (3×75 ml.), water (75 ml.) and saturatedaqueous sodium chloride solution (75 ml.), then dried (MgSO₄), filteredand evaporated in vacuo to yield a waxy solid that was used in the nextstep without further purification [2.0 g., 83% yield, ir (CHCl₃) 2250cm⁻¹ ].

D. Title Compound

Lithium aluminum hydride (800 mg., 21 mmoles) was added to a solution of1,3-di-O-(n-hexadecyl)-2-O-(3-cyanopropyl)-glycerol (2.0 g., 3.3 mmoles)in ether (100 ml.), and the mixture stirred for 60 hours at roomtemperature. Enough water to quench the reaction was added cautiously,followed by an additional 100 ml. of water. The resulting mixture wasstirred for another hour at room temperature and then extracted withether (3×100 ml.). The combined ether extract was washed with saturatedaqueous sodium chloride solution (3×75 ml.), dried (MgSO₄), filtered andevaporated in vacuo to an oil, which was purified by silica gelchromatography (elution with benzene:ethanol) and then dissolved inethanol. The solution was treated with hydrogen chloride gas and thenevaporated in vacuo to yield a solid, which was recrystallized fromethyl acetate [444 mg., 21% yield, m.p. 61.5°-63.5° C., n.m.r. (CDCl₃) δ3.67 (t, 2, --OCH₂ CH₂ CH₂ CH₂ NH₂), 3.55 (m, 9, --OCH[CH₂ OCH₂ C₁₅ H₃₁]₂), 3.10 (t, 2, --OCH₂ CH₂ CH₂ CH₂ NH₂), 1.50-2.00 (m, 4, --OCH₂ CH₂CH₂ CH₂ NH₂) and 0.80-1.50 (m, 62, aliphatic protons), elementalanalysis calculated: 72.23% C; 12.74% H; 2.16% N; found: 72.53% C;12.42% H; 2.10% N].

EXAMPLE 14 1,2-Di-O-(n-hexadecyl)-3-O-(3-aminomethylbenzyl)-glycerolHydrochloride A. 1,2-Di-O-(n-hexadecyl)-3-O-(3-cyanobenzyl)-glycerol

Sodium hydride (1.056 g. of 50 wt. % mineral oil dispersion, 22 mmoles)was added to a solution of 1,2-di-O-(n-hexadecyl)-glycerol (9.73 g., 18mmoles) in tetrahydrofuran (150 ml.) and the resulting solution stirredfor 20 minutes at room temperature under nitrogen. m-Cyanobenzyl bromide(4.0 g., 20 mmoles) was added and the reaction mixture stirred overnightat room temperature under nitrogen. Water (200 ml.) was then addedcautiously and the resulting mixture extracted with ethyl acetate (3×150ml.). The combined ethyl acetate extract was dried (MgSO₄), filtered andevaporated in vacuo to an oil (12 g.), which was purified by silica gelchromatography (elution with benzene:hexane) [8.0 g., 68% yield, oil, ir(CHCl₃) 2230 cm⁻¹ ].

B. Title Compound

A solution of 1,2-di-O-(n-hexadecyl)-3-O-(3-cyanobenzyl)-glycerol (1.0g., 1.5 mmoles) in ether (10 ml.) was slowly added under nitrogen to asuspension of lithium aluminum hydride (0.057 g., 1.5 mmoles) in ether(40 ml.), and the resulting mixtures stirred for one hour at refluxunder nitrogen and then cooled. Water (50 ml.) was added cautiously andthe mixture extracted with ether (3×50 ml.). The combined ether extractwas dried (MgSO₄), filtered and evaporated in vacuo to an oil, which waspurified by silica gel chromatography (elution with benzene:ethanol) andthen dissolved in ethyl acetate. The solution was treated with hydrogenchloride gas and then evaporated in vacuo to yield a solid, which wasrecrystallized from ethyl acetate [220 mg., 21% yield, m.p. 88°-90° C.,elemental analysis calculated: 74.14% C; 11.87% H; 2.01% N; found:74.35% C; 11.54% H; 2.15% N].

EXAMPLES 15-26

In like manner to that described in Example 14 the following compoundswere prepared by using the appropriate 1,3- or 1,2-di-O-(n-higher alkylor alkenyl)-glycerol and cyanobenzyl bromide as starting materials:

    __________________________________________________________________________     ##STR20##                                                 I                   ##STR21##                                                 II                                      Substitution                                                                  on                      Elemental Analysis               Ex.                                                                              Struc-            Phenyl                                                                              Molecular    M.P. Calculated                                                                             Found (%)               No.                                                                              ture                                                                              R.sub.1                                                                              R.sub.2                                                                              Ring  Formula      (°C.)                                                                       C  H  N  C  H  N                 __________________________________________________________________________    15 I   n-hexadecyl                                                                          n-hexadecyl                                                                          ortho C.sub.43 H.sub.81 O.sub.3 N .                                                              71-73                                                                              74.14                                                                            11.87                                                                            2.01                                                                             73.89                                                                            11.43                                                                            1.99              16 I   n-hexadecyl                                                                          n-hexadecyl                                                                          meta  C.sub.43 H.sub.81 O.sub.3 N .                                                              77-79                                                                              73.20                                                                            11.85                                                                            1.98                                                                             73.17                                                                            11.53                                                                            2.28              17 I   n-hexadecyl                                                                          n-hexadecyl                                                                          para  C.sub.43 H.sub.81 O.sub.3 N . HCl . H.sub.2                                                77-78                                                                              72.28                                                                            11.83                                                                            1.96                                                                             72.52                                                                            11.46                                                                            1.90              18 II  n-tetradecyl                                                                         n-tetradecyl                                                                         ortho C.sub.39 H.sub.73 O.sub.3 N . HCl . 1/4                                       H.sub.2 O    71-72                                                                              72.63                                                                            11.48                                                                            2.17                                                                             72.62                                                                            11.81                                                                            2.43              19 II  n-hexadecyl                                                                          n-hexadecyl                                                                          ortho C.sub.43 H.sub.81 O.sub.3 N .                                                              79-80                                                                              74.14                                                                            11.87                                                                            2.01                                                                             73.94                                                                            11.25                                                                            2.02              20 II  n-tetradecyl                                                                         n-tetradecyl                                                                         meta  C.sub.39 H.sub.73 O.sub.3 N .                                                              87-88                                                                              73.14                                                                            11.65                                                                            2.19                                                                             72.86                                                                            11.44                                                                            2.11              21 II  n-octadecyl                                                                          n-octadecyl                                                                          meta  C.sub.47 H.sub.89 O.sub.3 N .                                                              73-75                                                                              74.99                                                                            12.06                                                                            1.86                                                                             74.97                                                                            11.73                                                                            1.83              22 II  n-octadec-9-                                                                         n-octadec-9-                                                                         meta  C.sub.47 H.sub.85 O.sub.3 N . HCl . 1/2                                       H.sub.2 O    oil  74.50                                                                            11.57                                                                            1.85                                                                             74.40                                                                            11.08                                                                            2.08                     enyl   enyl                                                            23 II  n-tetradecyl                                                                         n-tetradecyl                                                                         para  C.sub.39 H.sub.73 O.sub.3 N .                                                              132-135                                                                            73.14                                                                            11.65                                                                            2.19                                                                             72.84                                                                            11.30                                                                            2.26              24 II  n-hexadecyl                                                                          n-hexadecyl                                                                          para  C.sub.43 H.sub.81 O.sub.3 N .                                                              117-119                                                                            74.14                                                                            11.87                                                                            2.01                                                                             74.33                                                                            11.55                                                                            2.15              25 II  n-octadecyl                                                                          n-octadecyl                                                                          para  C.sub.47 H.sub.89 O.sub.3 N .                                                              67-69                                                                              74.99                                                                            12.06                                                                            1.86                                                                             74.50                                                                            11.30                                                                            1.91              26 II  n-octadec-9-                                                                         n-octadec-9-                                                                         para  C.sub.47 H.sub.85 O.sub.3 N . HCl . 3/4                                       H.sub.2 O    oil  74.06                                                                            11.54                                                                            1.83                                                                             74.00                                                                            10.99                                                                            1.93                     enyl   enyl                                                            __________________________________________________________________________

EXAMPLE 27 1,2-Di-O-(n-hexadecyl)-3-O-(4-aminomethylphenyl)-glycerolHydrochloride A. 1,2-Di-O-(n-hexadecyl)-3-O-(p-tosyl)-glycerol

In like manner to that described in Example 13B the named compound wasprepared by reacting 1,2-di-O-(n-hexadecyl)-glycerol withp-toluenesulfonyl chloride. Purification was accomplished byrecrystallization from ethyl acetate [m.p. 53°-55° C., ir (CHCl₃) 1360and 1180 cm⁻¹ ].

B. 1,2-Di-O-(n-hexadecyl)-3-O-(4-cyanophenyl)-glycerol

A mixture of 1,2-di-O-(n-hexadecyl)-3-O-(p-tosyl)-glycerol (1.4 g., 2.0mmoles), sodium 4-cyanophenolate (0.5 g., 3.5 mmoles) and xylene (100ml.) was stirred for 16 hours at reflux. Since the reaction was not yetcomplete the xylene was removed by distillation and replaced byN,N-dimethylformamide (100 ml.), and the resulting solution stirred foranother 16 hours at 150° C. The reaction solution was then cooled,diluted with water (100 ml. and extracted with ether (2×100 ml.). Thecombined ether extract was washed consecutively with 3 N hydrochloricacid (100 ml.), 10 wt. % aqueous sodium bicarbonate solution (100 ml.)and water (100 ml.), dried (MgSO₄), filtered and evaporated in vacuo toan oil, which was purified by silica gel chromatography (elution withbenzene) [0.65 g., 50% yield, m.p. 53°-55° C., ir (CHCl₃) 2210 cm⁻¹ ].

C. Title Compound

1,2-Di-O-(n-hexadecyl)-3-O-(4-cyanophenol)-glycerol (0.60 g., 0.93mmole) was added to a suspension of lithium aluminum hydride (0.3 g.,7.9 mmoles) in ether (25 ml.), and the resulting mixture stirred for 30minutes at room temperature. Water (25 ml.) was then added cautiously,the ether and aqueous phases separated, and the latter extracted withether (3×25 ml.) and ethyl acetate (25 ml.). The five organic extractswere combined, dried (MgSO₄), filtered and evaporated in vacuo to anoil, which was dissolved in ether. The solution was treated withhydrogen chloride gas, causing precipitation of a solid [0.41 g., 64%yield, m.p. 110°-112° C., n.m.r. (CDCl₃) δ 4.02 (s, 2, --CH₂ NH₂),elemental analysis calculated: 73.91% C; 11.81% H; 2.05% N; found:73.62% C; 11.71% H; 2.14% N].

EXAMPLES 28-30

In like manner to that described in Example 27 B-C the followingcompounds were prepared from the aprpopriate tosylate (prepared as inExample 27A) and sodium cyanophenolate:

    __________________________________________________________________________     ##STR22##                                      I                              ##STR23##                                      II                                                             Elemental Analysis                           Example   Substitution on                                                                       Molecular      Calculated (%)                                                                         Found (%)                           Number                                                                             Structure                                                                          Phenyl Ring                                                                           Formula  M.P.(°C.)                                                                    C  H  N  C  H  N                             __________________________________________________________________________    28   I    meta    C.sub.42 H.sub.79 O.sub.3 N . HCl                                                      78-80 73.91                                                                            11.81                                                                            2.05                                                                             74.11                                                                            11.64                                                                            2.44                          29   I    para    C.sub.42 H.sub.79 O.sub.3 N . HCl                                                      120-122                                                                             73.91                                                                            11.81                                                                            2.05                                                                             73.94                                                                            11.37                                                                            2.04                          30   II   meta    C.sub.42 H.sub.79 O.sub.3 N . HCl                                                      84-86 73.91                                                                            11.81                                                                            2.05                                                                             74.00                                                                            11.34                                                                            2.04                          __________________________________________________________________________

EXAMPLE 31 1,2-Di-O-(n-hexadecyl)-3-O-[4-(3-aminopropyl)phenyl]-glycerolHydrochloride

In like manner to that described in Example 27 the named compound wasprepared by using sodium 4-(2-cyanoethyl)phenolate in place of sodium4-phenolate (m.p. 153°-155° C., elemental analysis calculated: 74.37% C;11.91% H; 1.97% N; found: 74.13% C; 11.44% H; 2.08% N).

EXAMPLE 32 1,2-Di(n-hexadecyloxy)-3-(3-aminomethylbenzylamino)-propaneDihydrochloride

1,2-Di-O-(n-hexadecyl)-3-O-(p-tosyl)-glycerol (3.48 g., 5.0 mmoles) wasadded to a solution of m-xylylenediamine (0.68 g., 5.0 mmoles) inN,N-dimethylformamide (20 ml.). The resulting mixture was stirred forone hour at 90° C. and then poured into ice water (150 ml.), causing theformation of solids which were isolated by filtration, purified bysilica gel chromatography (elution with benzene:ethanol) and thendissolved in ethyl acetate. The solution was treated with hydrogenchloride gas and then evaporated in vacuo to yield a solid, which wasrecrystallized from ethyl acetate [0.29 g., 8% yield, m.p. 78°-80° C.,n.m.r. (CDCl₃) δ 4.24 (s, 2, Ar--CH₂ NH--) and 4.37 (s, 2, Ar--CH₂ NH₂),elemental analysis calculated: 70.55% C; 11.57% H; 3.83% N; found:70.64% C; 11.29% H; 3.62% N].

EXAMPLE 331,2-Di-O-(n-hexadecyl)-3-O-(3-isopropylamino-2-hydroxypropyl)-glycerolHydrochloride A. 1,2-Di-O-(n-hexadecyl)-3-O-(2,3-epoxypropyl)-glycerol

A solution of 1,2-di-O-(n-hexadecyl)-3-allyl-glycerol (5.8 g., 10.0mmoles) and m-chloroperbenzoic acid (1.86 g., 10.8 mmoles) in benzene(50 ml.) was stirred at reflux for 16 hours. The reaction mixture wasthen cooled, treated with saturated aqueous sodium bisulfite solution(10 ml.) and saturated aqueous sodium bicarbonate solution (50 ml.), andextracted with ether (3×50 ml.). The combined ether extract was washedwith water (100 ml.), washed with saturated aqueous sodium chloridesolution (100 ml.), dried (MgSO₄), filtered and evaporated in vacuo toan oil (4.9 g., 82% yield, olefinic protons absent by n.m.r. analysis),which was purified by silica gel chromatography (elution withbenzene:ethyl acetate) (4.2 g., 70% yield, oil-solidified on standing).

B. Title Compound

A solution of 1,2-di-O-(n-hexadecyl)-3-O-(2,3-epoxypropyl)-glycerol (2.0g., 3.35 mmoles) in isopropylamine (40 ml.) was heated in a stainlesssteel bomb for 16 hours at 100° C., cooled, concentrated in vacuo anddissolved in ether (100 ml.). The ether solution was washed with 1 Nhydrochloric acid (100 ml.), dried (MgSO₄), filtered, treated withcharcoal, filtered again, and then cooled by immersion of the flask in aDry Ice-acetone bath, causing formation of a precipitate. Theprecipitate was isolated by filtration (1.3 g.) and purified by silicagel chromatography (elution with benzene:ethanol) [720 mg., solidcontained about 1/2 mole H₂ O per mole named product, 31% yield, m.p.55°-57° C., n.m.r. (CDCl₃) δ 1.45 (d, 6, --NHCH[CH₃ ]₂), elementalanalysis calculated: 70.19% C; 12.50% H; 2.00% N; found: 70.10% C;12.19% H; 1.87% N].

EXAMPLES 34-37

In like manner to that described in Example 33B the following compoundswere prepared by reacting the appropriate 2,3-epoxide (prepared as inExample 33A) and alkylamine:

    __________________________________________________________________________     ##STR24##                                          I                          ##STR25##                                          II                                                            Elemental Analysis                        Example          Molecular          Calculated (%)                                                                         Found (%)                        Number                                                                             Structure                                                                          R.sub.3                                                                              Formula      M.P. (°C.)                                                                   C  H  N  C  H  N                          __________________________________________________________________________    34   I    CH.sub.2 CH.sub.3                                                                    C.sub.40 H.sub.83 O.sub.4 N . HCl                                                          53-54 68.97                                                                            12.44                                                                            2.01                                                                             69.35                                                                            12.08                                                                            1.94                       35   I    CH(CH.sub.3).sub.2                                                                   C.sub.41 H.sub.85 O.sub.4 N . HCl                                                          67-68 71.10                                                                            12.52                                                                            2.02                                                                             70.54                                                                            12.18                                                                            2.05                       36   I    C(CH.sub.3).sub.3                                                                    C.sub.42 H.sub.87 O.sub.4 N . HCl                                                          60-61 71.39                                                                            12.55                                                                            1.98                                                                             71.55                                                                            12.35                                                                            1.79                       37     II C(CH.sub.3).sub.3                                                                    C.sub.42 H.sub.87 O.sub.4 N . HCl . 1/2 H.sub.2 O                                          50-51 70.49                                                                            12.53                                                                            1.96                                                                             70.66                                                                            12.31                                                                            1.83                       __________________________________________________________________________

EXAMPLE 38 1,2-Di-O-(n-hexadecyl)-3-O-(3-amino-2-hydroxypropyl)-glycerolHydrochloride A.1,2-Di-O-(n-hexadecyl)-3-O-(3-azido-2-hydroxypropyl)-glycerol

A solution of sodium azide (0.5 g., 7.7 mmoles) in water (5 ml.) wasadded to a refluxing solution of1,2-di-O-(n-hexadecyl)-3-O-(2,3-epoxypropyl)-glycerol (3.3 g., 5.5mmoles) in 1,4-dioxane (100 ml.), and the resulting solution stirred atreflux for 16 hours. Since the reaction was not yet complete, additionalsodium azide (0.5 g., 7.7 mmoles) was added and the reaction stirred atreflux for another 16 hours. The reaction solution was then cooled,concentrated in vacuo, diluted with water (100 ml.) and extracted withether (3×100 ml.). The combined ether extract was washed with water (100ml.), dried (MgSO₄), filtered and evaporated in vacuo to an oil whichsolidified on standing [2.2 g., 62% yield, ir (CHCl₃) 2105 cm⁻¹ ].

B. Title Compound

Lithium aluminum hydride (300 mg., 7.9 mmoles) was added to a solutionof 1,2-di-O-(n-hexadecyl)-3-O-(3-azido-2-hydroxypropyl)-glycerol (2.2g., 3.4 mmoles) in ether (100 ml.), and the resulting mixture stirredfor one hour at room temperature. Ethanol (5 ml.) and water (200 ml.)were added to quench the reaction, and the mixture then extracted withether (2×100 ml.). The combined ether extract was dried (MgSO₄),filtered and evaporated in vacuo. The resulting product was purified bysilica gel chromatography (elution with benzene:ethanol) and thenconverted to the hydrochloride salt [800 mg., solid contained about 2moles H₂ O per mole named product, 34% yield, m.p. 149°-150° C., n.m.r.(CDCl₃) δ 4.00-4.35 (m, 1, --OCH₂ CHOCHCH₂ NH₂), 3.33-3.73 (m, 11, C₁₅H₃₁ CH₂ OCH₂ CH[OCH₂ C₁₅ H₃₁ ]CH₂ OCH₂ --), 3.03-3.25 (m, 2, --OCH₂CHOHCH₂ NH₂) and 0.87-1.67 (m, 62, aliphatic protons), elementalanalysis calculated: 66.48% C; 12.33% H; 2.04% N; found: 66.68% C;11.85% H;2.02% N].

EXAMPLE 39 1,3-Di-O-(n-hexadecyl)-2-O-(3-amino-2-hydroxypropyl)-glycerol

In like manner to that described in Example 38 the named compound wasprepared from 1,3-di-O-(n-hexadecyl)-2-O-(2,3-epoxypropyl)-glycerol(prepared as in Example 33A) (free base, m.p. 61°-63° C., elementalanalysis calculated: 74.33% C; 12.96% H; 2.28% N; found: 74.49% C;13.10% H; 2.12% N).

EXAMPLE 40 1,2-Di-O-(n-hexadecyl)-3-O-(2-aminopropyl)-glycerolHydrochloride A.1,2-Di-O-(n-hexadecyl)-3-O-[2-(p-tosyloxy)propyl]-glycerol

In like manner to that described in Example 13A and B,1,2-di-O-(n-hexadecyl)-3-O-allyl-glycerol was reacted with BMS, and theresulting 2-hydroxypropyl and 3-hydroxypropyl compounds converted totheir corresponding tosylates. A separation was not attempted at thisstage; the mixture of tosylates was used directly in the next step.

B. 1,2-Di-O-(n-hexadecyl)-3-O-(2-azidopropyl)-glycerol

The resulting mixture of tosylates (3.0 g., 4.0 mmoles) was dissolved inN,N-dimethylacetamide (50 ml.) and treated with a solution of sodiumazide (0.326 g., 5.0 mmoles) in water (5 ml.) for 16 hours at 90° C. Thereaction solution was then cooled, diluted with water (200 ml.), andextracted with ether (2×150 ml.). The combined ether extract was washedwith water, dried (MgSO₄), filtered, and evaporated in vacuo to an oil[2 g., 81% yield, ir (CHCl₃) 2100 cm⁻¹ ], a mixture of the 2-azidopropyland 3-azidopropyl compounds, which was used without further purificationin the next step.

C. Title Compound

The resulting mixture of azides (2 g., 3.2 mmoles) was dissolved inether (100 ml.), treated with lithium aluminum hydride (0.4 g., 10.5mmoles), and allowed to stir for 2 hours at room temperature. Excesshydride was destroyed by cautious addition of ethanol (10 ml.) and water(150 ml.), and the mixture then extracted with ether (2×100 ml.). Thecombined ether extract was dried (MgSO₄), filtered, and concentrated invacuo to an oil (1.8 g.), which was purified by silica gelchromatography (elution with benzene:ethanol) and then converted to thehydrochloride salt by dissolution and treatment with hydrogen chloridegas. The salt was recrystallized from ethyl acetate (0.21 g., solidcontained about 1/2 mole H₂ O per mole named product, 10% yield, m.p.56°-58° C. elemental analysis calculated: 71.03% C; 12.70% H; 28.1% N;found: 71.11% C; 12.91% H; 2.16% N).

EXAMPLE 41 1,2-Di-O-(n-octadecyl)-3-O-(2-aminopropyl)-glycerolHydrochloride

In like manner to that described in Example 40A,1,2-di-O-(n-octadecyl)-3-O-(2-hydroxypropyl)-glycerol was prepared from1,2-di-O-(n-octadecyl)-3-O-allyl-glycerol. The named compound wasprepared from 1,2-di-O-(n-octadecyl)-3-O-(2-hydroxypropyl)glycerol inlike manner to that described in Example 40 B-C (solid contained about 1mole H₂ O per mole named product, m.p. 65°-67° C., elemental analysiscalculated: 71.19% C; 12.80% H; 1.98% N; found: 71.12% C; 12.52% H;1.92% N).

EXAMPLE 42 1,2-Di(n-hexadecyloxy)-3-aminopropane Hydrochloride

In like manner to that described in Example 40B,1,2-di-O-(n-hexadecyl)-3-O-(p-tosyl)-glycerol was converted to1,2-di(n-hexadecyloxy)-3-azidopropane. This intermediate was convertedto the title compound in like manner to that described in Example 40C(m.p. 78°-80° C., elemental analysis calculated: 72.93% C; 12.94% H;2.43% N; found: 73.08% C; 13.08% H; 2.65% N).

EXAMPLE 43 1,3-Di(n-hexadecyloxy)-2-aminopropane Hydrochloride

In like manner to that described in Example 42 the named compound wasprepared from 1,3-di-O-(n-hexadecyl)-2-O-(p-tosyl)-glycerol (prepared asin Example 27A) (m.p. 58°-60° C., elemental analysis calculated: 72.93%C; 12.94% H; 2.43% N; found: 72.65% C; 13.02% H; 2.59% N).

EXAMPLE 44 1,2-Di(n-hexadecyloxy)-4-aminobutane Hydrochloride

In like manner to that described in Example 42 the named compound wasprepared by using sodium cyanide in place of sodium azide (m.p. 86°-87°C., elemental analysis calculated: 73.25% C; 12.97% H; 2.37% N; found:73.52% C; 12.64% H; 2.50% N).

EXAMPLE 45 1,3-Di(n-hexadecyloxy)-2-(3-aminopropylamino)propaneDihydrochloride A. 1,3-Di(n-hexadecyloxy)-2-(2-cyanoethylamino)propane

A mixture of 1,3-di(n-hexadecyloxy)-2-aminopropane (500 mg., 0.93mmoles), acrylonitrile (75 ml.) and 2 wt. % aqueous sodium hydroxidesolution (75 ml.) was heated to 60° C. Tetrabutyl ammonium hydroxide (1ml. of 40 wt. % aqueous solution) was then added and the resultingmixture stirred for 15 minutes at 90° C. The reaction mixture was thencooled, causing precipitation of solids, which were isolated byfiltration and found (TLC) to contain a large quantity of unreactedstarting material. Using fresh acrylonitrile and aqueous sodiumhydroxide solution in each cycle, the solids were treated two more timesby the above procedure. The third cycle solid product was purified bysilica gel chromatography (elution with toluene:ethyl acetate) [200 mg.,36% yield, m.p. 45°-46° C., ir (CHCl₃) 2250 cm⁻¹, n.m.r. (CDCl₃) δ 3.07(t, 2, --NHCH₂ CH₂ CN) and 2.53 (t, 2, --NHCH₂ CH₂ CN)].

B. Title Compound

A mixture of 1,3-di(n-hexadecyloxy)-2-(2-cyanoethylamino)-propane (200mg., 0.34 mmoles), tetrahydrofuran (10 ml.), ethanol (20 ml.) and Raneynickel catalyst (0.2 g.) was saturated with ammonia gas and thenhydrogenated (50 psi) for about 4 hours at room temperature. Thereaction mixture was then filtered and evaporated in vacuo to an oil,which was purified by silica gel chromatography (elution withtoluene:ethyl acetate: ethanol: methanol) and then dissolved in ethylacetate. The solution was treated with hydrogen chloride gas, causingprecipitation of solids [10 mg., solid contained about 2.5 moles H₂ Oper mole named product, 4% yield, m.p. 235°-236° C., elemental analysiscalculated: 63.65% C; 12.51% H; 3.90% N; found: 63.60% C; 11.84% H;3.75% N].

EXAMPLE 46 1,2-Di-O-(n-hexadecyl)-3-O-(4-amidinophenyl)-glycerolHydrochloride

A solution of 1,2-di-O-(n-hexadecyl)-3-O-(4-cyanophenyl)-glycerol (3.5g., 5.45 mmoles), ethanol (10 ml.) and 1,4-dioxane (100 ml.) wassaturated with hydrogen chloride gas at 0° C., and allowed to react for16 hours at ambient temperature. The reaction solution was thenevaporated in vacuo to an oil, the oil dissolved in ethanol (100 ml.),and the resulting solution saturated with ammonia gas, stirred for 3hours at reflux, diluted with water (150 ml.), evaporated in vacuo toremove the majority of the ethanol, and extracted with chloroform (3×150ml.). The combined chloroform extract was dried (MgSO₄), filtered andevaporated in vacuo to yield a solid, which was purified by silica gelchromatography (elution with benzene:ethanol) and then dissolved inethyl acetate. The solution was treated with hydrogen chloride gas andthen evaporated in vacuo to yield a solid, which was recrystallized fromethyl acetate [1.0 g., 26% yield, m.p. 220°-222° C., ir (CHCl₃) 1670cm⁻¹, elemental analysis calculated: 72.53% C; 11.45% H; 4.03% N; found:72.67% C; 11.38% H; 4.12% N].

EXAMPLE 47 1,2-Di-O-(n-hexadecyl)-3-O-(3-amidinobenzyl)-glycerolHydrochloride

The named compound was prepared from1,2-di-O-(n-hexadecyl)-3-O-(3-cyanobenzyl)-glycerol in like manner tothat described in Example 46 [solid contained about 2 moles H₂ O permole named product, 20% yield, m.p. 155°-157° C., ir (CHCl₃) 1670 cm⁻¹,elemental analysis calculated: 69.27% C; 11.48% H; 3.76% N; found:69.11% C; 10.63% H; 3.83% N].

EXAMPLE 481,2-Di-O-(n-hexadecyl)-3-O-[3-(1-hydroxy-2-t-butylaminoethyl)-benzyl]-glycerolHydrochloride A. 1,2-Di-O-(n-hexadecyl)-3-O-(3-formylbenzyl)-glycerol

A solution of 1,2-di-O-(n-hexadecyl)-3-O-(3-cyanobenzyl)-glycerol (5.0g., 7.6 mmoles) and diisobutylaluminum hydride (1.17 g., 8.2 mmoles) inbenzene (25 ml.) was stirred for 16 hours at ambient temperature. Thereaction mixture was treated with methanol (4.22 ml.) and water (2.5ml.) and stirred to decompose unreacted hydride, and then filtered andextracted with benzene (3×25 ml.). The combined benzene extract wasdried (Na₂ SO₄), filtered and evaporated in vacuo to an oil, which waspurified by silica gel chromatography (elution with benzene) [2.0 g.,40% yield, oil, ir (CHCl₃) 1700 cm⁻¹, n.m.r. (CDCl₃) δ 10.1 (s, 1,--ArCHO)].

B. 1,2-Di-O-(n-hexadecyl)-3-O-[3-(1,2-epoxyethyl)benzyl]-glycerol

A suspension of sodium hydride (3.23 g. of a 57 wt. % dispersion inmineral oil, 67 mmoles) in dimethylsulfoxide (117 ml.) was heated undera nitrogen atmosphere at 70° to 75° C. until hydrogen evolution stopped(45 min.). Tetrahydrofuran (88 ml.) was added and the mixture cooled to0° to 5° C. Trimethylsulfonium iodide (13.67 g., 67 mmoles) was thenadded in portions, followed by rapid addition of a solution of1,2-di-O-(n-hexadecyl)-3-O-(3-formylbenzyl)-glycerol (7.0 g., 10.6mmoles) in tetrahydrofuran (58 ml.). The resulting mixture was stirredfor 16 hours at room temperature, poured into water (200 ml.) andextracted with ether (3×180 ml.). The combined ether extract was washedwith water (2×100 ml.) and saturated aqueous sodium chloride solution(100 ml.), dried (MgSO₄), filtered and evaporated in vacuo to an oil(7.0 g., 98% yield), which was sufficiently pure to be used in the nextstep.

C. Title Compound

A mixture of t-butylamine (30 ml.) and1,2-di-O-(n-hexadecyl)-3-O-[3-(1,2-epoxyethyl)-benzyl]-glycerol (2.0 g.,3.0 mmoles) was heated for 9 hours at 100° C. in a steel bomb. Thereaction mixture was cooled, t-butylamine removed by evaporation invacuo, and the resulting oil purified by silica gel chromatography(elution with benzene:ethanol) and then dissolved. The solution wassaturated with hydrogen chloride gas and then evaporated in vacuo toyield a solid, which was recrystallized from ethyl acetate [630 mg.,solid contained about 1 mole H₂ O per mole named product, 27% yield,m.p. 49°-51° C., n.m.r. (CDCl₃) δ 1.47 (s, 9, --C[CH₃ ]₃), elementalanalysis calculated: 71.99% C; 11.83% H; 1.75% N; found: 71.86% C;11.30% H; 1.69% N].

EXAMPLE 491,3-Di-O-(n-hexadecyl)-2-O-[3-(1-hydroxy-2-t-butylaminoethyl)-benzyl]-glycerolHydrochloride

In like manner to that described in Example 48 A-B,1,3-di-O-(n-hexadecyl)-2-O-(3-cyanobenzyl)-glycerol (prepared as inExample 14A) was converted to1,3-di-O-(n-hexadecyl)-2-O-[3-(1,2-epoxyethyl)-benzyl]-glycerol. Thetitle compound was prepared by reacting said epoxy compound witht-butylamine in like manner to that described in Example 48C (solidcontained about 1 mole H₂ O per mole named product, m.p. 43°-45° C.,elemental analysis calculated: 71.99% C; 11.83% H; 1.75% N; found:72.06% C; 11.43% H; 1.71% N).

EXAMPLE 501,2-Di-O-(n-hexadecyl)-3-O-[3-(1-hydroxy-2-isopropylaminoethyl)-benzyl]-glycerolHydrochloride

In like manner to that described in Example 48C the named compound wasprepared by using isopropylamine in place of t-butylamine (solidcontained about 3/4 mole H₂ O per mole named product, m.p. 53°-55° C.,elemental analysis calculated: 72.17% C; 11.79% H; 1.79% N; found:72.11% C; 11.55% H; 1.92% N).

EXAMPLE 511-[2,3-Di(n-hexadecyloxy)propyl]-4-aminomethyl-4-phenylpiperidineDihydrochloride A.1-[2,3-Di(n-hexadecyloxy)propyl]-4-cyano-4-phenylpiperidine

A mixture of 1,2-di-O-(n-hexadecyl)-3-O-(p-tosyl)-glycerol (6.96 g., 10mmoles), 4-cyano-4-phenylpiperidine hydrochloride (2.23 g., 10 mmoles),triethylamine (2 ml.) and N,N-dimethylformamide (40 ml.) was stirred for16 hours at 95° to 100° C. The reaction mixture was then cooled, dilutedwith water (200 ml.) and extracted with ethyl acetate (3×150 ml.). Thecombined ethyl acetate extract was dried (MgSO₄), filtered andevaporated in vacuo to an oil (6 g.), which was purified by columnchromatography (elution with benzene:ethyl acetate) [oil, ir (CHCl₃)2220 cm⁻¹ ].

B. Title Compound

A solution of1-[2,3-di(n-hexadecyloxy)propyl]-4-cyano-4-phenylpiperidine (2.5 g., 3.6mmoles) in ether (100 ml.) was treated with lithium aluminum hydride(0.4 g., 10.5 mmoles), and the resulting mixture stirred for 4 hours atroom temperature. The reaction mixture was treated cautiously with water(100 ml.) and extracted with ether (3×100 ml.). The combined etherextract was dried (MgSO₄), filtered and evaporated in vacuo to an oil,which was purified by silica gel chromatography (elution withbenzene:ethanol) and then dissolved. The solution was treated withhydrogen chloride gas and then evaporated in vacuo to yield a solid,which was recrystallized from ethyl acetate (1.1 g., solid containedabout 3/4 mole H₂ O per mole named product, 40% yield, m.p. 132°-134°C., elemental analysis calculated: 70.60% C; 11.53% H; 3.50% N; found:70.74% C; 11.34% H; 3.40% N).

EXAMPLES 52-54

In like manner to that described in Example 51 the following compoundswere prepared from the appropriate 1,2-di-O-(n-alkyl oralkenyl)-3-O-(p-tosyl)-glycerol (prepared as in Example 27A):

    __________________________________________________________________________     ##STR26##                                                                                                  Elemental Analysis                              Example       Molecular       Calculated (%)                                                                         Found (%)                              Number                                                                             R        Formula   M.P.(°C.)                                                                    C  H  N  C  H  N                                __________________________________________________________________________    52   n-tetradecyl                                                                           C.sub.43 H.sub.80 O.sub.2 N.sub.2 . 2HCl                                                140-142                                                                             69.46                                                                            11.25                                                                            3.75                                                                             69.45                                                                            11.00                                                                            3.45                             53   n-octadecyl                                                                            C.sub.51 H.sub.96 O.sub.2 N.sub.2 . 2HCl                                                115-117                                                                             71.95                                                                            11.60                                                                            3.29                                                                             71.67                                                                            11.19                                                                            3.38                             54   n-octadec-9-enyl                                                                       C.sub.51 H.sub.92 O.sub.2 N.sub.2 . 2HCl                                                118-120                                                                             71.17                                                                            11.23                                                                            3.25                                                                             71.06                                                                            10.84                                                                            3.09                             __________________________________________________________________________

EXAMPLE 561,2-Di-O-(n-hexadecyl)-3-O-(2-[di(2-hydroxyethyl)amino]ethyl)-glycerolHydrochloride

In like manner to that described in Example 11 the named compound wasprepared by reacting 1,2-di-O-(n-hexadecyl)-3-O-formylmethyl-glycerolwith di(2-hydroxyethyl)amine (solid contained about 1/4 mole H₂ O permole named product, m.p. 194°-195° C., elemental analysis calculated:69.06% C; 12.20% H; 1.96% N; found: 69.12% C; 11.76% H; 1.90% N).

EXAMPLE 57 In Vivo Activity of1,3-Di-O-(n-hexadecyl)-2-O-(3-aminopropyl)-glycerol HydrochlorideAgainst EMC Virus

Formulation as an emulsion was accomplished by melting and mixing equalparts of the named compound, polysorbate 80, and glycerin, and thendispersing the mixture in hot water under vigorous mixing. Theformulation was then adjusted to final concentrations of 0.14 M sodiumchloride and 0.01 M sodium phosphate, pH 7. Further dilutions were madewith 0.14 M sodium chloride -0.01 M sodium phosphate, pH 7 buffersolution.

Three groups of ten female albino mice (20-25 g. body weight) were given0.5 ml. intraperitoneal injections containing dosage levels of 1.5, 5and 15 mg. of the named compound/kg. body weight, respectively. A fourthcontrol group of ten mice was given no such injection. Eighteen totwenty-four hours later all four groups were challenged with 0.2 ml.subcutaneous injection containing 20 times the LD₅₀, the dosage levelcausing a 50% death rate in unprotected mice in ten days, ofencephalomyocarditis (EMC) virus. Survival data were recorded over thenext ten days and the relative survival (S_(r)) calculated:

    ______________________________________                                        Dosage Level of                                                               Named Compound                                                                              S.sub.r (average of seven experiments)                          ______________________________________                                        15     mg./kg.    61                                                          5                 45                                                          1.5               24                                                          ______________________________________                                    

Antiviral activity is expressed as the relative survival (S_(r)) inexperimental groups compared to the control on the tenth day afterchallenge. S_(r) is defined by the formula ##EQU1## wherein S_(r)=relative survival

S_(x) =percent survival after ten days in experimental group

x_(i) =number of survivors on the ith day in experimental group

e_(i) =number of survivors on the ith day in control group

EXAMPLES 58-86

In like manner to that described in Example 57 the in vivo activityagainst EMC virus was determined for the compounds listed below.

    ______________________________________                                               Compound                                                               Example                                                                              Prepared in   S.sub.r at Dosage Level (mg./kg.) of                     Number Example Number                                                                              15      5     1.5   0.5                                  ______________________________________                                        58     14            71      49    20    6                                    59     15            62      47    18    --                                   60     16            80      72    20    --                                   61     17            74      46    4     --                                   62     18            57      42    3     --                                   63     19            64      58    15    --                                   64     20            74      44    34    --                                   65     21            46       7    6     --                                   66     22            45       5    4     --                                   67     23            70      36    3     --                                   68     24            37      31    12    0                                    69     25            44      30    8     --                                   70     26            68      43    2     --                                   71     27            66      45    17    --                                   72     28            68      28    33    --                                   73     29            60      63    16    --                                   74     30            53      34    16    7                                    75     31            56      35    20    6                                    76     32            63      41    31    --                                   77     42            33      26    26    --                                   78     43            30      30    7     --                                   79     44            51      20    7     --                                   80     46            32       3    0     --                                   81     47            56      16    6     --                                   82     51            77      58    26    --                                   83     52            76      85    42    --                                   84     53            52      42    32    --                                   85     54            79      72    28    --                                   86     49            99      26    54    --                                   ______________________________________                                    

EXAMPLE 87 Reduction of Virus Yield on Human Polyp Cells In Vitro by1,3-Di-O-(n-hexadecyl)-2-O-(3-aminopropyl)-glycerol Hydrochloride

Growth medium was prepared by supplementing Eagle's minimum essentialmedium (100 ml.) with 100× concentrated antibioticantimycotic solution(2 ml.), 200 mM glutamine solution (1 ml.), 100× concentratednonessential amino acids solution (1 ml.), 100 mM sodium pyruvatesolution (1 ml.), and heat-inactivated fetal calf serum (10%). Each wellof 96-well microtiter plates was seeded with about 50,000 human nasalpolyp cells suspended in 0.2 ml. growth medium. The plates were thenincubated for 8-10 days at 37° C. in a 5% CO₂ atmosphere to establishmonolayers of cells.

At the end of the 8-10 day cell growth period confluent monolayers onthe plates were washed four times with phosphate buffered saline andimmediately afterward treated with 0.2 ml. per well of maintenancemedium containing 10, 5.0, 1.0, 0.5, 0.1 and 0 μg./ml. of the namedcompound, respectively. The maintenance medium was identical to thegrowth medium described above except that the level of said fetal calfserum was 2%. The plates were incubated for another 18 hours at 37° C.,and the monolayers then washed four times with phosphate buffered salineto remove the named compound, challenged with a composition containingabout 1000 times the TCID₅₀, i.e. the dosage level causing a 50%infection rate in unprotected cultures, of vesicular stomatitis virus(VSV) for a two hour (37° C.) adsorption period, washed four times withphosphate buffered saline to remove unadsorbed virus particles, andrefed with 0.2 ml. per well of said maintenance medium. The plates werethen incubated for 7 hours at 37° C., and the culture fluid from 5-8replicate cells harvested from each plate, stored frozen in test tubes,and then titrated for the amount of infectious virus present inmicrotiter plates of L-929 mouse fibroblasts. The L-929 mouse cultureswere scored microscopically and analyzed about three to four days later,with the following percentage decreases in virus yield (with respect tothe control) determined for the five concentrations of named compoundtested:

    ______________________________________                                        Percentage Reduction of Virus Yield                                           Concentration (μg./ml.) of Named Compound                                  10      5.0       1.0       0.5     0.1                                       ______________________________________                                        94%     90%       84%       75%     <68%                                      ______________________________________                                    

EXAMPLES 88-121

In like manner to that described in Example 87 the reduction of virusyield on human polyp cells in vitro was determined for the compoundslisted below.

    __________________________________________________________________________                     Percentage Reduction of Virus Yield.sup.a                    Example                                                                            Compound Prepared in                                                                      Concentration (μg./ml.)                                   Number                                                                             Example Number                                                                            10   5.0 1.0 0.5  0.1                                        __________________________________________________________________________    88    6          +    +   -   -    ND                                         89    9          ND   ±                                                                              -   -    ND                                         90   11          +    +   -   ND   ND                                         91   12          +    +   -   ND   ND                                         92   13          +    ±                                                                              -   ND   ND                                         93   14          +    +   -   -    -                                          94   18          +    -   -   ND   ND                                         95   19          +    +   -   -    -                                          96   20          +    ±                                                                              -   -    -                                          97   22          +    ±                                                                              -   -    -                                          98   23          +    ±                                                                              -   -    -                                          99   28          +    +   -   -    -                                          100  30          +    +   -   ND   ND                                         101  31          +    +   -   ND   ND                                         102  33          +    +   -   ND   ND                                         103  34          ND   +   -   ND   ND                                         104  35          +    ±                                                                              -   ND   ND                                         105  36          +    -   -   ND   ND                                         106  37          +    +   -   ND   ND                                         107  38          +    +   -   -    ND                                         108  40          +    +   -   ND   ND                                         109  41          ND   +   -   ND   ND                                         110  42          +    +   -   -    ND                                         111  44          +    +   -   ND   ND                                         112  45          +    -   -   ND   ND                                         113  47          +    +   -   -    -                                          114  56          +    -   -   ND   ND                                         115  48          +    +   -   -    ND                                         116  49          -    -   -   -    ND                                         117  50          +    +   -   -    ND                                         118  51          +    ±                                                                              -   -    ND                                         119  52          +    +   -   -    ND                                         120  53          +    +   -   -    ND                                         121  54          +    -   -   -    ND                                         __________________________________________________________________________     .sup.a + .tbd. >68% reduction;                                                ± .tbd. ˜ 68% reduction;                                             - .tbd. <68% reduction;                                                       ND .tbd. not done.                                                       

EXAMPLE 122 Ability of1,3-Di-O-(n-hexadecyl)-2-O-(3-aminopropyl)-glycerol Hydrochloride toInduce Circulating Interferon

A mixture of equal weights of the named compound, polysorbate 80 andglycerol was fused and then homogenized in hot 0.14 M sodium chloridecontaining 0.01 M sodium phosphate, pH 7 (PBS). The resultingoil-in-water emulsion was readily diluted with PBS for administration.

Female Swiss mice (20-25 g. body weight) were injected (0.5 ml.,intraperitoneal) with a quantity of the above diluted emulsioncontaining 25 mg. of the named compound/kg. body weight. Eight, twelve,sixteen and twenty hours after injection samples of plasma werewithdrawn from four mice and pooled. Serial dilutions in L-15(Leibovitz) medium containing 5% fetal calf serum were incubated inmicrotiter plates overnight at 37° C. on confluent monolayers of L-929mouse fibroblasts. The monolayers were then washed with protein-freemedium, challenged with 10 times the TCID₅₀, i.e. the dosage levelcausing a 50% infection rate in unprotected cultures, of vesicularstomatitis virus (VSV) for a one hour (37° C.) adsorption period,washed, retreated with L-15 medium containing 5% fetal calf serum andthen incubated again for 48 hours at 37° C. The L-929 cultures were thenscored microscopically for viral cytopathology and analyzed, with theplasma interferon level, the reciprocal of the plasma dilutionconferring 50% protection to the L-929 monolayers, determined.

A second experiment followed the above procedure, except that the micewere injected with 10 mg. of the named compound/kg. body weight andsamples of peritoneal wash were taken from four mice and pooled at six,nine, twelve, fifteen and eighteen hours after injection. The sampleswere taken by exposing the peritoneal membrane, injecting 1 ml. ofHank's balanced salt solution containing 100 penicillin units/ml. and100 μg. streptomycin/ml. into the peritoneal cavity, briefly massagingthe abdomen, and then aspirating the peritoneal wash.

The following data were obtained from these two experiments:

    ______________________________________                                                Interferon Levels (units/ml.)                                         Source of                                                                             Time (hrs.) after Injection                                           Interferon                                                                            6       8      9    12   15   16   18   20                            ______________________________________                                        Plasma  --      34     --    67  --   52   --   40                            Peritoneal                                                                    wash    <16     --     768  320  448  --   448  --                            ______________________________________                                    

EXAMPLES 123-129

In like manner to that described in Example 122 the ability to inducecirculating interferon was determined for the compounds listed below.

    __________________________________________________________________________                     Interferon Levels (units/ml.).sup.a                                                          Interferon Levels (units/ml.).sup.b           Example                                                                            Compound Prepared in                                                                      Time (hrs.) after Injection                                                                  Time (hrs.) after Injection                   Number                                                                             Example Number                                                                            8   12  16  20 6  9  12 15 18                                __________________________________________________________________________    123  14          <20 23  75  90 <13                                                                              <13                                                                              39 35 72                                124  15           20 71  54  68 <18                                                                              <18                                                                              43 59 32                                125  16          <18 138 217 163                                                                              <13                                                                              <13                                                                              43 92 57                                126  24          <20 28  60  70 <13                                                                              <13                                                                              91 109                                                                              50                                127  32           38 33  45  35 <13                                                                               79                                                                              49 52 <13                               128  47          <17 19  45  99 <16                                                                               32                                                                              96 640                                                                              512                               129  51           70.sup.c                                                                         100  120.sup.d                                                                         100.sup.e                                                                       <13                                                                              109                                                                              284                                                                              312                                                                              224                               __________________________________________________________________________     .sup.a Interferon source  plasma                                              .sup.b Interferon source  peritoneal wash                                     .sup.c <20 units/ml. at 6 hrs., 70 units/ml. at 9 hrs.                        .sup.d 15 hrs.                                                                .sup.e 18 hrs.                                                           

EXAMPLE 130 Enhancement of Polyinosinic-Polycytidylic Acid [Poly(I:C)]-induced Cellular Resistance to Viral Infection by1,3-Di-O-(n-hexadecyl)-2-O-(3-aminopropyl)-glycerol Hydrochloride

Growth medium was prepared by supplementing Eagle's minimum essentialmedium (100 ml.) with 100× concentrated antibiotic-antimycotic solution(2 ml.), 200 mM glutamine solution (1 ml.), and heat-inactivated fetalcalf serum (5%). Mouse L-929 fibroblasts were suspended in growthmedium, and each well of 96-well microtiter plates was seeded with 0.2ml. of said suspension containing 20,000 to 30,000 cells. The plateswere incubated for 2 to 4 days at 37° C. in a 5% CO₂ atmosphere toestablish monolayers of cells. The plates were washed four times withphosphate buffered saline immediately prior to treatment.

Poly (I:C) was prepared at concentrations of 5.0, 1.0, 0.2 and 0.04μg./ml. in the medium described above minus calf serum. 0.1 ml. of eachdilution was combined in a checkerboard arrangement on the L-929 cellmonolayers with 0.1 ml. dilutions containing 20.0, 4.0, 0.8, 0.16 and0.032 μg. of the named compound per ml. said serum-free medium. Controlwells were exposed to either poly(I:C) or the named compound alone. Theplates were incubated for 6 hours at 37° C. in a 5% CO₂ atmosphere,washed four times with phosphate buffered saline, and refed with 0.1 ml.per well growth medium containing 2% fetal calf serum. After 18 morehours of incubation, the plates were scored for toxicity and thenchallenged with 0.1 ml. per well of a vesicular stomatitis virus (VSV)suspension containing 10 to 30 times the TCID₅₀ (tissue cultureinfective dose causing a 50% infection rate). The plates were incubatedfor another 3 to 4 days and then scored microscopically forcytopathogenic effect (CPE). Cells protected from virus infection werefree of CPE. The minimum protective dose (MPD) of poly (I:C) alone wasnoted, and the amount of enhanced or augmented antiviral activity causedby combination with the named compound recorded for each dilution levelof said named compound.

    ______________________________________                                        Enhancement of Poly (I:C)-induced                                             Cellular Resistance to Viral Infection                                        Concentration (μg./ml.) of Named Compound                                  20.0     4.0       0.8       0.16    0.032                                    ______________________________________                                        125X     125X      125X      5X      <5X                                      ______________________________________                                    

Note: combining poly(I:C) with named compound provides some antiviraleffect as increasing poly (I:C) concentration indicated number of times.

EXAMPLES 131-162

In like manner to that described in Example 130 the enhancement ofpoly(I:C)-induced cellular resistance to viral infection was determinedfor the compounds listed below.

    ______________________________________                                                                 Poly (I:C) Enhancement.sup.a                         Example Compound Prepared in                                                                           Concentration (μg./ml.)                           Number  Example Number   20      4.0   0.8                                    ______________________________________                                        131      6               +       +     -                                      132      7               +       +     -                                      133     14               +       +     ±                                   134     15               +       +     ±                                   135     16               +       +     ±                                   136     17               +       +     ±                                   137     18               +       +     ±                                   138     19               +       +     ±                                   139     20               +       +     ±                                   140     21               +       ±  -                                      141     22               -       -     -                                      142     23               +       +     ±                                   143     24               +       +     ±                                   144     25               +       +     -                                      145     26               +       -     -                                      146     27               +       +     -                                      147     28               +       +     -                                      148     29               +       +     -                                      149     30               +       +     ±                                   150     31               +       +     -                                      151     32               +       +     ±                                   152     40               +       +     ±                                   153     41               +       +     -                                      154     42               +       +     +                                      155     43               +       +     +                                      156     44               +       +     ±                                   157     46               +       +     ±                                   158     47               +       +     ±                                   159     51               +       +     ±                                   160     52               +       +     -                                      161     53               +       +     ±                                   162     54               ±    -     -                                      ______________________________________                                         .sup.a + .tbd. >5X enhancement;                                               ± .tbd. ˜5X enhancement;                                             - .tbd. <5X enhancement;                                                      ND .tbd. not done.                                                       

What is claimed is:
 1. A compound selected from those of the formula##STR27## and the pharmaceutically acceptable acid addition saltsthereof, wherein R₁ and R₂ are each selected from the group consistingof normal alkyl of from 12 to 20 carbon atoms and normal alkenyl nothaving a double bond in the 1-position of from 12 to 20 carbon atoms. 2.A compound of claim 1 wherein R₁ and R₂ are each normal alkyl of from 14to 18 carbon atoms.
 3. A compound of claim 2 wherein R₁ and R₂ eachcontain the same number of carbon atoms.
 4. The compound of claim 1wherein R₁ and R₂ are each n-tetradecyl.
 5. The compound of claim 1wherein R₁ and R₂ are each n-hexadecyl.
 6. The compound of claim 1wherein R₁ and R₂ are each n-octadecyl.
 7. A pharmaceutical compositioncontaining an antivirally effective amount of a compound of claim 1 asthe essential active ingredient in a pharmaceutically acceptablecarrier.
 8. A method of prophylactically controlling a viral infectionin a mammal which comprises administering an amount effective toprophylactically control said viral infection of a compound of claim 1.9. A method of inducing the production of interferon in a mammal whichcomprises administering an amount effective to induce the production ofinterferon of a compound of claim 1.